Corum:DNA Hybridization: Difference between revisions

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==Procedure==
==Procedure==
# Combine sense and antisense oligonucleotides. Vortex and spin.
# Combine 5 μL sense oligonucleotide and 5 μL antisense oligonucleotide. Vortex and spin.
# Double seal tube airtight with paraffin and press firmly in weighted holder.
# Double seal tube airtight with parafilm and press firmly in weighted holder.
# Boil 800 mL water in a 1L beaker.
# Boil 800 mL water in a 1L beaker.
# Cool on bench 3 min to ~95 °C.
# Wait 1 min.
# Place weighted holder with sealed oligonucleotides into the beaker.
# Place weighted holder with sealed oligonucleotides into the beaker.
# Incubate on bench overnight or until water is about room temperature (~4 hr). At the end of this process, the complimentary ssDNA's are hybridized into as single dsDNA linker with sticky ends at 1mM final concentration.
# Incubate on bench overnight or until water is about room temperature (~4 hr). At the end of this process, the complimentary ssDNA's are hybridized into as single dsDNA linker with sticky ends at 1mM final concentration.
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